• Korean Journal of Microbiology
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• v.21 no.4
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• pp.221-228
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• 1983

An improved procedure for the rapid purification of glucose-6-phosphate dehydrogenase from extracts of Saccharomyces cerevisiae was developed by using affinity chromatography. Among six affinty media tested, $NADP^+ -agarose$ and Affi-gel Blue were more effective than others (i.e., Affi-gel Red, AMP-agarose, ATP-agarose, and $NAD^+ -agarose$). Conditions to desorb the enzyme bound to the affinity media were examined to increase the purity as well as yield. The best result was obtained when the column was developed with a linear gradient of KCl (0-1.0M). In case of Affi-gel Blue, introduction of $NAD^+$ (15mM) washing step prior to the salt gradient was most effective to remove $NAD^+ -binding$ proteins. For a large scale preparation of G-6-P dehydrogenase higher recovery was obtained by Affi-gel Blue than $NADP^+ -agarose$, however, the purity of the enzyme was decreased by 10 times if the former was used as the affinity medium. The capacity of Affi-gel Blue for G-6-P dehydrogenase was found to be 5 times higher than that of $NADP^+ -agarose$. Furthermore Affi-gel Blue could be reused repeatedly and its preparation is relatively easier and less expensive than $NADP^+ -agarose$.

• Journal of Life Science
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• v.27 no.1
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• pp.57-66
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• 2017

Traditional Korean fermented herbal plants are potential sources of new food that promote health, but they are still produced by yeast, fungi or bacteria fermentation. In the present work, mushroom (Paecilomyces tenuipes and Cordyceps militaris) fungal dongchunghacho were used to fermented Glycyrrhiza uralensis Fischer (licorice) or mixed with pupa. The pupa were tested as solid substrates for the production of corcycepin, liquiritin, and liquiritigenin. The fermented substrates were analyzed the content of cordycepin, liquiritin, liquiritigenin, and glycirrhizin productivity and inhibitory activity of NO. The cordycepin content of 70% EtOH extract from the fermented mixture of licorice and 50% pupa with C. militaris increased maximum at 33 times. Pupa was very excellent for the production of cordycepin. The liquiritin content was decreased in all the assays inoculated with P. tenuipes and C. militaris dongchunghachos. The liquiritigenin content was higher when fermented with P. tenuipes than C. militaris. The addition of pupa significantly reduced the liquiritin content and glycyrrhizin production. As a result, the liquiritigenin content increased in fermented P. tenuipes and C. militaris, and liquiritin and glycyrrhizin decreased. The inhibition of NO production in the different ethanolic extracts fermented with licorice and pupa was also significantly increased and higher than that of a nonfermented extract in higher polar solvent extracts. The contents of cordycepin and biological active compounds were altered in accordance with the concentration of pupa and fungi. This study provides basic data for use in developing dongchunghacho fungi as a functional food resource.

• Korean Journal of Microbiology
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• v.50 no.4
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• pp.351-359
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• 2014

A beverage was produced by the fermentation of mixed extracts from the various fruits, vegetables, algae, and medical herbs. The physicochemical properties of the fermented beverage of plant extracts (FBPE) and microbial diversity were analyzed in cultures enriched from FBPE using 16S and 26S rRNA gene sequence analyses. The pH, acidity, $^{\circ}brix$, reducing sugar, and alcohol contents of the FBPE were determined to be the 3.48, 1.68%, 70.0, 1,026 g/L, and 3.5%, respectively. The most abundant free sugar and organic acid in the FBPE were glucose (567.83 g/L) and tartaric acid (93.68 mg/L), respectively. Lactobacillus homohiochii was the predominant species in all enriched culture samples: 100% of the species in 0B (0% sugar) and 40B (40% sugar) libraries and 95.6% of 20B library (20% sugar). Lactobacillus fructivorans was detected in the 20B library. The predominant species in the samples of enrichment cultures collected from FBPE with three different sugar concentrations were: Candida zeylanoides (45.2%) in the 0Y library (0% sugar), Candida lactis-condensi (35.7%) and C. zeylanoides (35.7%) in the 20Y library (20% sugar), and C. lactis-condensi (38.1%) in the 40Y library (40% sugar). This result may provide a useful frame of reference for further analyses of microbial population dynamics in FBPE.

• The Korean Journal of Mycology
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• v.4 no.1
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• pp.31-44
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• 1976

The studies were carried out to examine the effects of supplementation of nutritional substances and physical conditions in substrate on the mycelial growth and yield of fresh sporophores of winter mushroom, Flammulina velutipes(Curt. ex Fr.) Sing. and to obtain further informations on the nutritional requirements of the fungus with reference to improvement of substrate through [analysis of chemical composition of the substrates during the cultivation period. The results obtained are summarized as follows: 1. The best yield of fresh sporophores, 84.4 g per 280 g substrate in a bottle, was obtained from the mixture of poplar sawdust 10 and rice bran 3 by volume when Flammulina velutipes was cultivated on the poplar sawdust supplemented by rice bran, wheat bran, cattle manure and various combinations of these materials as nutrient sources. The substrates of poplar sawdust 10 plus rice bran 3 and 2 or wheat bran 3 with a higher yield of fresh sporophores showed a comparatively higher content of total nitrogen. total sugar, and potassium. 2. The mycelial growth of the fungus was compared on the substrates of poplar sawdust supplemented by the several nutrient sources and poplar sawdust alone. The fastest linear growth occurred on substrates of poplar sawdust alone and poplar sawdust plus cattle manure deficient in sugar and nitrogen sources, but mycelial density was more sparse on the substrates. Also, growth in a solution extracted from these substrates was very meager. 3. In the substrates which varied with bulk density and moisture content optimum bulk density and moisture content for mycelial growth was 0.2g/cc and 72% on a dry weight basis, respectively, but the highest yield of fresh sporophores was obtained at the bulk density of 0.3g/cc and moisture content of 67%. 4. By increasing the ratio of rice bran in poplar sawdust the loss of total weight and ash, content at each stage was increased, and during the cultivation period of 75 days, loss of total weight of the substrates at inoculation was 17.8 to 28.8% and ash content increased about 12%. 5. 11 to 14% of the cellulose and 3 to 4% of the lignin content per original substrate were decreased without a great difference depending of the mixing ratio of rice bran. The soluble glucose concentration in the substrates was increased during the same period. 6. In the process of vegetative and reproductive growth of the fungus upon the substrates, the total nitrogen was increased in quantity per dry weight of sample but was reduced in absolute quantity to a minute extent. There is no great changes in content of organic nitrogen including amino acid nitrogen, and hydrolysable ammonium nitrogen during the vegetative growth period, but occurrence of sporophores resulted in a decrease in the nitrogen content of these forms. On the one hand, by an increase of additive amounts of rice bran, nitrogen contents of these forms were higher and the reduction range during the reproductive growth period became wider. 7. Mycelial growth of the fungus was accelerated in various liquid media supplemented with organic nitrogen sources such as peptone and yeast extract in comparison with addition of inorganic nitrogen sources. Furthermore, mycelial growth was mere vigorous in the media with higher content of organic nitrogen sources.

• Applied Biological Chemistry
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• v.30 no.1
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• pp.31-39
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• 1987

This study has been investigated the effect of Ganordema lucidum extract on Saccharomyces cerevisiae growth and physiology. Sacch. cerevisiae was inoculated in Hayduck solution medium which were added 0, 0.1, 0.5, 1.0% extracts of G. lucidum and fermented at $30^{\circ}C$ for 5 days respectively. Some results about cell number, alcohol content and carbon dioxide products during fermentation are as follows: $CO_2$ evolution of yeasts by addition of extract of G. lucidum was more increased than control after the fermentation for 120 hours. It was the most abundant by addition of 1.0% extract of pot-culture G. lucidum. The cell number of yeasts during the fermentation w as more increased than control by addition of extract of G. lucidum. It was by addition of extract of pot-culture G. lucidum that the cell number of yeasts was more increased than by each addition of extract of wood-culture G. lucidum and G. lucidum. Dry weight of yeasts was systematically increased in addition of extract of pot 0.5%>pot 1.0%>wild 1.0%>wood 1.0%=wood 0.5%>wild 0.5%>wild 0.1%>pot 0.1%>wood 0.1%>control in order. It was by addition of extract of pot-culture G. lucidum that. the dry weight of yeasts was more increased than by addition of woodculture G. lucidum and wild G. lucidum. Alcohol quantity by addition of extract of G. lucidum was increased more than 3 times after the fermentation for 72 hours compared with control but there was no any difference among them after the fermentation for 120 hours. The rate of sugar-consumption and fermentation of yeast by addition of extract of G. lucidum was highly increased during the early fermentation. As times went, there was no difference among them during the subsequent fermentation.

• The Korean Journal of Mycology
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• v.40 no.4
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• pp.244-253
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• 2012

The effect of fermented Rhus verniciflua stem bark (FRVSB) extract on the microbial count, enzyme activity, concentrations of free amino acids and organic acids, and physiochemical properties of doenjang (soybean paste) was evaluated during brine fermentation. The FRVSB extract increased the total free amino acid concentration by 1.3-3.1-fold on the $42^{nd}$ day of brine fermentation. After the filtration of brine, the following microbial counts were obtained in the doenjang: bacteria, $0.3{\times}10^8-12.0{\times}10^8$ cfu/g; mold, $3.0{\times}10^4-21.0{\times}10^4$ cfu/g; yeast, $1.0{\times}10^4-2.0{\times}10^4$ cfu/g; Escherichia coli, not detected; and Bacillus cereus, $3.0{\times}10^2-25.0{\times}10^2$ cfu/g. The FRVSB extract addition enhanced the protein and starch degrading activity by 13.8-26.0% and 16.1-35.1%, respectively. The extract increased the total free amino acid content by 1.4-3.0-fold. Lactic acid, acetic acid, and pyroglutamic acid were the predominant organic acids in doenjang. Moreover, the proximate composition, pH, moisture, ash, salt, and amino nitrogen content were increased.

• Korean Journal of Medicinal Crop Science
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• v.11 no.1
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• pp.53-61
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• 2003

Cirsium japonicum var. ussuriense were extracted with methanol and then fractionated with nhexane, EtOAc and BuOH to get active fractions. And their antioxidant and antimicrobial activities in each fraction were determined. Ethyl acetate and butanol fraction of Cirsium japonicum var. ussuriense showed strong antioxidant activities, but hexane fraction did not show any activities. But in the antimicrobial test, Ethyl acetate fraction showed strong antimicrobial activities except to Aspergillus awamori, Asperigillus niger. Especially, Ethyl acetate fraction showed the strongest activities against Bacillus subtilis. And aqueous fraction showed the strongest activities against Cladosporium herbarum, Hypocrea nigricans. This study was performed to determine the antimutagenic and cytotoxic effect of Cirsium japonicum var. ussuriense methanol extract on Salmonella typhimurium TA98, TA100 and cancer cell lines using Ames test and cytotoxicity assay, respectively. Cancer cell lines include human lung carcinoma(A549), human breast adenocarcinoma(MCF-7) and human hepatocellular carcinoma (Hep3B). Futher fractionations with hexane, ethyl acetate, butanol and water from methanol extract of Cirsium japonicum var. ussuriense were performed to obtain effective fraction, methanol extract showed 60.14% inhibition effect on the mutagenesis induced by MNNG against TA100, while 77% and 72.5% inhibition was observed on the mutagenesis induced by 4NQO against TA98 and TA100, respectively. and methanol extract showed 82.25% and 73.7% inhibitory effect on the mutagenesis induced by Trp-P-1 against TA98 and TA100, respectively. methanol extract showed the strongest effect against A549, MCF-7 and Hep3B at the same concentration compared to those of other fration.

• KSBB Journal
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• v.18 no.1
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• pp.55-61
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• 2003

To isolate and purify the antimicrobial and antitumor agents in Xanthium strumarium L. hydrothermal extract. The crude extract was extracted in ether or ethylacetate under neutral, acidic, and alkali conditions. The antimicrobial activity of each extract was tested against 16 strains of bacteria, 2 strains of yeast, and 2 strains of fungus. The ether neutral extract (XE-N) exhibited the strongest growth inhibition upon the 8 strains of gram-positive bacteria, 6 strains of gram-negative bacteria and Cryptococcus neoformans. Fluorescein diacetate (FDA) testing of XE-N and XEA-N showed growth inhibition of the 3 strains of E. coli, S. aureus and C. albicans even at 30 ng/mL, with the exception of p. aeruginosa. XE-N-S1 and XE-N-S3 from neutral ether extract (XE-N), XE-N-S3 from the acidic ether extract (XE-A), and XEA-N-S1 from ethylacetate (XEA-N) were purified as antimicrobial and antitumor agents. However all purified compounds decomposed with the exception of XE-N-S1. The results upon the antitumor activities of the crude extract and of its purified compounds, showed that XE-N-S1 had the best antitumor activity against HeLa cells. In terms of antitumor activity against HepG2 cells, XE-N-S1 and XE-N-S3 were superior, and against HT29 cells XE-N and XE-N-Sl were good, against Saos2, NCI H522, NCI H1703, Clone M3 cells XE-N-51 was very good, and against LN CAP cells XE-N-S3 was the best. Comparing of cellular toxicities various extracts and purified compounds with the existing antitumor agents, XE-A, XEA-A and XEA-B had the lowest toxicity, and XE-B had a lower toxicity than etoposide. XE-N-S1 and XE-N-S3 showed higher toxicities than etoposide, and the toxicity of XE-A-S3 was higher than that of etoposide, and lower than that of csplatin.

• Journal of Life Science
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• v.29 no.3
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• pp.369-375
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• 2019

Microtubules form through the polymerization of ${\alpha}-$ and ${\beta}-tubulin$, and tubulin transport plays an important role in defining the rate of microtubule growth inside cellular appendages, such as the cilia and flagella. Heterotrimeric kinesin 2 is a molecular motor member of the kinesin superfamily (KIF) that moves along the microtubules to transport multiple cargoes. It consists of two motor subunits (KIF3A and KIF3B) and a kinesin-associated protein 3 (KAP3), forming a heterotrimeric complex. Heterotrimeric kinesin 2 interacts with many different binding proteins through the cargo-binding domains of the KIF3s, but these binding proteins have not yet been specified. To identify these proteins for KIF3A, we performed yeast two-hybrid (Y2H) screening and found a specific interaction with ${\beta}2-tubulin$ (Tubb2), a microtubule component. Tubb2 was found to bind to the cargo-binding domain of KIF3A but did not interact with KIF3B, KIF5B, or kinesin light chain 1 in the Y2H assay. The carboxyl-terminal region of Tubb2 is essential for interaction with KIF3A. Other Tubb isoforms, including Tubb1, Tubb3, Tubb4, and Tubb5, also interacted with KIF3A in the Y2H screening. However, ${\alpha}1-tubulin$ (Tuba1) did not interact with KIF3A. In addition, an antibody to KIF3A specifically co-immunoprecipitated the KIF3B and KAP3 associated with Tubb2 from mouse brain extracts. In combination, these results suggest that a heterotrimeric kinesin 2 motor protein is capable of binding to tubulin and may transport it in cells.